Dissociation kinetics of actinomycin D from individual GpC sites in DNA
Dissociation kinetics of actinomycin D from individual GpC sites in DNA
We have examined the kinetics of dissociation of actinomycin from GpC sites in several DNA fragments containing synthetic DNA inserts, by a variation of the footprinting technique. Complexes of the ligand with radiolabelled DNA fragments were dissociated by adding a large excess of unlabelled calf thymus DNA. Samples were removed from this mixture at subsequent time intervals and subjected to DNase I footprinting. The rate of disappearance of the footprints varied considerably between the GpC sites located in different sequence environments. Actinomycin dissociates more slowly from GpC sites flanked by (AT)n than An · Tn. Within regions of alternating AT, TGCA represents a better binding site than AGCT, and CGCA is a better binding site than GGCA. GpC sites flanked by (AC)n · (GT)n present good binding sites; in this context, dissociation from CGCG is faster than from TGCA.
Actinomycin, Dissociation, Footprinting, Kinetics
164-170
Fletcher, Michael C.
81b0f822-354c-4e8a-a3da-e0ead19c1e90
Fox, Keith R.
9da5debc-4e45-473e-ab8c-550d1104659f
1996
Fletcher, Michael C.
81b0f822-354c-4e8a-a3da-e0ead19c1e90
Fox, Keith R.
9da5debc-4e45-473e-ab8c-550d1104659f
Fletcher, Michael C. and Fox, Keith R.
(1996)
Dissociation kinetics of actinomycin D from individual GpC sites in DNA.
European Journal of Biochemistry, 237 (1), .
(doi:10.1111/j.1432-1033.1996.0164n.x).
Abstract
We have examined the kinetics of dissociation of actinomycin from GpC sites in several DNA fragments containing synthetic DNA inserts, by a variation of the footprinting technique. Complexes of the ligand with radiolabelled DNA fragments were dissociated by adding a large excess of unlabelled calf thymus DNA. Samples were removed from this mixture at subsequent time intervals and subjected to DNase I footprinting. The rate of disappearance of the footprints varied considerably between the GpC sites located in different sequence environments. Actinomycin dissociates more slowly from GpC sites flanked by (AT)n than An · Tn. Within regions of alternating AT, TGCA represents a better binding site than AGCT, and CGCA is a better binding site than GGCA. GpC sites flanked by (AC)n · (GT)n present good binding sites; in this context, dissociation from CGCG is faster than from TGCA.
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Published date: 1996
Keywords:
Actinomycin, Dissociation, Footprinting, Kinetics
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Local EPrints ID: 475871
URI: http://eprints.soton.ac.uk/id/eprint/475871
ISSN: 0014-2956
PURE UUID: 766d879d-d0af-48e3-8bfc-f95808a8d6ff
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Date deposited: 29 Mar 2023 16:47
Last modified: 17 Mar 2024 02:34
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Author:
Michael C. Fletcher
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